To raise this antibody bovine intermediate filaments were prepared from spinal cords by the method of Delacourte et al. and the cytoskeletal material was dissolved in 6M urea. To ensure greater specificity for NFL, animals were boosted with recombinant mouse NFL purified from bacteria.
This antibody was generated in chicken by standard procedures and immunoglobulin was extracted from egg yolk. This is the chicken homologue of mammalian IgG and can be used in the same general way, with the caveat that this type of antibody does not bind either Protein A or Protein G.

Previously labelled as 68kDa Neurofilament.

Properties

Form

Liquid

Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.

Storage buffer

Preservative: 0.065% Sodium azideConstituent: PBS

Concentration information loading...

Purification notes

The IgY preparation was made by chloroform delipidation of egg yolk followed by polyethylene glycol precipitation.

Primary antibody notes

To raise this antibody bovine intermediate filaments were prepared from spinal cords by the method of Delacourte et al. and the cytoskeletal material was dissolved in 6M urea. To ensure greater specificity for NFL, animals were boosted with recombinant mouse NFL purified from bacteria.
This antibody was generated in chicken by standard procedures and immunoglobulin was extracted from egg yolk. This is the chicken homologue of mammalian IgG and can be used in the same general way, with the caveat that this type of antibody does not bind either Protein A or Protein G.

Associated products

Applications

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application

Abreviews

Notes

IHC

1/2000.

ICC/IF

1/5000.

WB

1/10000. Predicted molecular weight: 63 kDa.

Using chemiluminescence, 1/10000 or lower.

Target

Function

Neurofilaments usually contain three intermediate filament proteins: L, M, and H which are involved in the maintenance of neuronal caliber.

Involvement in disease

Defects in NEFL are the cause of Charcot-Marie-Tooth disease type 1F (CMT1F) [MIM:607734]. CMT1F is a form of Charcot-Marie-Tooth disease, the most common inherited disorder of the peripheral nervous system. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathy or CMT1, and primary peripheral axonal neuropathy or CMT2. Neuropathies of the CMT1 group are characterized by severely reduced nerve conduction velocities (less than 38 m/sec), segmental demyelination and remyelination with onion bulb formations on nerve biopsy, slowly progressive distal muscle atrophy and weakness, absent deep tendon reflexes, and hollow feet. CMT1F is characterized by onset in infancy or childhood (range 1 to 13 years).Defects in NEFL are the cause of Charcot-Marie-Tooth disease type 2E (CMT2E) [MIM:607684]. CMT2E is an autosomal dominant form of Charcot-Marie-Tooth disease type 2. Neuropathies of the CMT2 group are characterized by signs of axonal regeneration in the absence of obvious myelin alterations, normal or slightly reduced nerve conduction velocities, and progressive distal muscle weakness and atrophy.

Sequence similarities

Belongs to the intermediate filament family.

Domain

The extra mass and high charge density that distinguish the neurofilament proteins from all other intermediate filament proteins are due to the tailpiece extensions. This region may form a charged scaffolding structure suitable for interaction with other neuronal components or ions.

Post-translationalmodifications

O-glycosylated.Phosphorylated in the Head and Rod regions by the PKC kinase PKN1, leading to inhibit polymerization.

ICC/IF image of ab24520 stained SKNSH cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab24520, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was ab96947, DyLight® 488 goat anti-chicken IgY (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM